EP0608636B1 - Cycloisomaltooligosaccharide; Enzym und Verfahren zu ihrer Herstellung, und Verfahren zur Herstellung von dem Enzym - Google Patents

Cycloisomaltooligosaccharide; Enzym und Verfahren zu ihrer Herstellung, und Verfahren zur Herstellung von dem Enzym Download PDF

Info

Publication number
EP0608636B1
EP0608636B1 EP93310579A EP93310579A EP0608636B1 EP 0608636 B1 EP0608636 B1 EP 0608636B1 EP 93310579 A EP93310579 A EP 93310579A EP 93310579 A EP93310579 A EP 93310579A EP 0608636 B1 EP0608636 B1 EP 0608636B1
Authority
EP
European Patent Office
Prior art keywords
cycloisomaltooligosaccharide
producing
enzyme
cycloisomaltooligosaccharides
dextran
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
EP93310579A
Other languages
English (en)
French (fr)
Other versions
EP0608636A1 (de
Inventor
Tetsuya C/O Noda Institute For Ogumam
Tatsuo C/O Noda Institute For Horiuchi
Koichiro C/O Noda Institute For Tobe
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kikkoman Corp
Noda Institute for Scientific Research
Original Assignee
Kikkoman Corp
Noda Institute for Scientific Research
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from JP34944492A external-priority patent/JP3075873B2/ja
Priority claimed from JP05153456A external-priority patent/JP3117328B2/ja
Application filed by Kikkoman Corp, Noda Institute for Scientific Research filed Critical Kikkoman Corp
Publication of EP0608636A1 publication Critical patent/EP0608636A1/de
Application granted granted Critical
Publication of EP0608636B1 publication Critical patent/EP0608636B1/de
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
    • C08B37/0021—Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins

Definitions

  • the present invention relates to isomaltooligosaccharides, isomaltooligosaccharide synthase forming said oligosaccharides from dextran, and a process for producing said oligosaccharides by using said enzyme or a microorganism capable of producing said enzyme.
  • a cyclodextrin of 6-8 glucose residues bound by ⁇ -1,4 linkages is well known among cycloligosaccharides.
  • the cyclodextrin owing to its hydrophobicity in the center of the ring, possesses the ability (inclusion ability) to bind to a variety of hydrophobic substances.
  • the compound has been applied and developed for stabilization and improvements in solubility etc. of various substance in a wide range of fields such as medicaments, cosmetics, foods, etc. (Handbook of Amylases and Related Enzymes, pp. 233-243 (1988), compiled by Japanese Amylase Research Society.
  • cyclooligosaccharides such as cyclosophoraose composed of 17-24 glucose residues in ⁇ -1,2 linkage, cyclogentiooligosaccharides of 3-4 glucose residues in ⁇ -1,6 linkage, cycloawaodorin with 6 rhamnose molecules cyclized, cyclomannohexaose with 6 mannose molecules cyclized, etc., have been enzymatically or chemically synthesized.
  • the object of the present invention is to provide cycloisomaltooligosaccharides of glucoses bound by ⁇ -1,6 linkages expected highly useful, a process for producing the same, cycloisomaltooligosaccharide synthase forming said oligosaccharides from dextran, and a process for producing said enzyme.
  • the present invention encompasses:
  • the present cyclooligosaccharides differ completely from conventional cyclooligosaccharides in respect of the properties, and are completely new in that they consist of glucoses bound by ⁇ -1,6 linkages. Owing to their cyclic structure, the present compounds have an inclusion ability useful for stabilization, and solubilization of substances of different size not covered by cyclodextrin, and they are used specifically as inclusion agents for medicaments and foods.
  • the present cycloisomaltooligosaccharide synthase has the enzymatic chemical properties of:
  • the present enzyme differs in the properties from conventional cyclooligosaccharide synthases, cyclodextrin glucanotransferase (referred to hereinafter as “CGTase”; EC 2.4.1.19), cycloinulooligosaccharide synthase (referred to hereinafter as “CFTase”) and cyclic ⁇ -1,2 glucan synthetase in that it forms from dextran a cyclooligosaccharide of glucoses bound by ⁇ -1,6 linkages.
  • CCTase cyclodextrin glucanotransferase
  • CFTase cycloinulooligosaccharide synthase
  • cyclic ⁇ -1,2 glucan synthetase cyclic ⁇ -1,2 glucan synthetase in that it forms from dextran a cyclooligosaccharide of glucoses bound by ⁇ -1,6 linkages
  • the present enzyme is a multifunctional enzyme which, besides intramolecular transglucosylation reaction, catalyzes the reaction (coupling reaction) of forming an isomaltooligosaccharide having an OH acceptor bound thereto by intermolecular transglucosylation reaction of a cycloisomaltooligosaccharide and a suitable OH acceptor, as well as the reaction (disproportionation reaction) of forming isomaltooligosaccharides of various polymerization degrees from isomaltooligosaccharides.
  • the microorganism used in the present invention may be any of the microorganisms belonging to the genus Bacillus capable of forming cycloisomaltooligosaccharide synthase, and an example is Bacillus sp. T-3040.
  • Bacillus sp. T-3040 is a wild strain obtained by screening from soil, and possesses the following bacterial properties:
  • the bacterial strain T-3040 is a spore-forming Gram-positive microorganism, and the present inventors have identified it as a microorganism belonging to the genus Bacillus , but they could not identified its species in spite of extensive identification on the basis of Bergey's Manual of Systematic Bacteriology, vol. 2.
  • This microorganism identified merely as a bacterial strain belonging to the genus Bacillus , is designated Bacillus sp. T-3040 and has been deposite with Fermentation Research Institute, Agency of Industrial Science and Technology, Japan under FERM BP-4132.
  • the microorganism according to the present invention is aerobically cultured in the same manner as for conventional microorganisms, usually shake culture in liquid medium, spinner culture under aeration, etc.
  • the medium used is composed e.g. of a suitable nitrogen source (e.g., casein hydrolyzates such as peptone, polypeptone, bactotrypton, etc., or soy bean protein hydrolyzates such as soytone etc.), carbon source (e.g. sugar contents such as glucose, glycerin, etc.), and as necessary, yeast extract, vitamins such as riboflavin etc. and phosphates, magnesium salt, sodium chloride, minerals e.g. trace metals, and dextran etc. serving as starting materials for the present cycloisomaltooligosaccharides.
  • the pH of the medium may be in any pH range in which the present microorganism can grow, preferably at pH 6-8.
  • the microorganism is cultured e.g. under shaking or stirring under aeration, usually at 20-40 °C, preferably 30 °C and for 16 hours-6 days, preferably 3 days.
  • the present cyclooligosaccharide synthase is obtained from the resulting culture, for example, according to the following enzyme purification step.
  • a microorganism-free enzyme preparation produced merely by centrifuging the culture and then concentrating the resulting supernatant through a membrane can be used for the production of the present cyclooligosaccharides , and as necessary, a purified standard is obtained in a usual manner from this crude enzyme preparation.
  • a cyclooligosaccharide synthase standard of high purity can be obtained by treating a microorganism-free culture as described below.
  • the present enzyme is allowed to act on dextran for production of cycloisomaltooligosaccharides.
  • the enzyme reaction is effected at 10-60 °C, preferably 40 °C, at pH 4.5-8.0, preferably 5.0-6.5 and for 10-72 hours, preferably 48 hours.
  • the reaction solution may be stirred and an organic solvent such as methanol, ethanol, etc., may be added to the reaction solution.
  • the cycloisomaltooligosaccharides can be formed by use of the aforementioned microorganism-free concentrate of the culture in the same efficiency as attained by use of the purified enzyme.
  • Any purification method for conventional oligosaccharides can also be employed as the means of obtaining the present oligosaccharides from the reaction solution. Methods known to the art, such as cooling, treatment with organic solvent or active carbon, chromatography on an active carbon column or cyclooligosaccharide-specific adsorption column, etc., may be used singly or in combination.
  • the cycloisomaltooligosaccharides of the present invention can also be obtained by e.g. the recovering step as described below from a culture of said microorganism in a medium containing ⁇ -1,6 glucan such as dextran.
  • the culture is centrifuged for removal of the microorganisms, and then concentrated through a membrane.
  • the microorganism-free concentrate thus obtained is subjected to a purification method for conventional cyclodextrin, to give high-purity cyclooligosaccharide fractions.
  • a high-purity sample of each cyclooligosaccharide can be obtained by a purification means such as high performance liquid chromatography (HPLC) on a partition-adsorption column etc.
  • HPLC high performance liquid chromatography
  • Conventional methods such as cooling, treatment with organic solvent or activated charcoal, chromatography on an activated charcoal column or cyclooligosaccharide-specific adsorption column, etc., may be used singly or in combination for purification of the cycloisomaltooligosacchrides from the microorganism-free culture concentrate.
  • novel cycloisomaltooligosaccharides composed of glucose residues bound by ⁇ -1,6 linkage useful as inclusion agents for medicaments, foods, etc., and a process for producing said oligosaccharides of high purity in high yield by using a microorganism belonging to the genus Bacillus .
  • cycloisomaltooligosaccharise synthase allowing said cycloisomaltooligosaccharides to be efficiently produced in high yield in a very simple procedure.
  • the present invention is extremely useful in industry.
  • FIG. 1 shows infrared absorption spectra of the 3 cycloisomaltooligosaccharides of the present invention.
  • FIG. 2 shows the result of enzymatic analysis of the 3 cycloisomaltooligosaccharides of the present invention.
  • FIG. 3 shows the result of analysis in HPLC of the 3 cycloisomaltooligosaccharides of the present invention (A: an eluate in chromatography on activated charcoal, B-D: separated cycloligosaccharides).
  • FIG. 4 shows profiles in HPLC of the 3 cycloisomaltooligosaccharides of the present invention.
  • a liquid medium (tap water, pH 7.0) composed of 1 % dextran T2000, 1 % peptone, 0.5 % NaCl and 0.1 % yeast extract was put in 15 ml test tube and then sterilized at 120 °C for 20 min.
  • the culture solution 3 ml
  • the culture solution 3 ml
  • the microorganism was aerobically cultured at 30 °C, 0.25 vvm, and 350 rpm for 2 days under shaking.
  • the culture was centrifuged at 8000 rpm for 20 min., to separate into a bacterial precipitate and a supernatant as a microorganism-free culture.
  • This crude cycloisomaltooligosaccharide solution was concentrated in a rotary evaporator and then subjected to HPLC on YMC PA43 column (a preparative partition-adsorption chromatography column produced by Yamamura Kagaku Co., Ltd.), thus separating into each cycloisomaltooligosaccharide.
  • each fraction was concentrated in a rotary evaporator, and the resulting concentrate was analyzed for oligosaccharide purity in HPLC on TSK gel Amide 80 column (a partition-adsorption chromatography column produced by Tohso Co., Ltd.). The result is shown in FIG. 3 B-D. A 98 % or more purity was attained for each oligosaccharide.
  • the oligosaccharide fractions were lyophilized to give about 60 mg cycloisomaltoheptaose, about 200 mg cycloisomaltooctaose and about 100 mg cycloisomaltononaose, respectively.
  • a liquid medium (tap water, pH 7.0) composed of 1 % dextran 40 (produced by Meito Sangyo Co., Ltd.), 1 % peptone (Kyokuto Seiyaku Kogyo Co., Ltd.), 0.5 % NaCl and 0.1 % yeast extract (produced by Difco), were sterilized at 120 °C for 20 min.
  • the bacterial strain, Bacillus sp. T-3040 (FERM BP-4132), was inoculated onto the medium and cultured at 30 °C for 1 day under shaking.
  • 1000 ml of the culture thus obtained was inoculated onto 300 l medium in a 500 l tank with the same medium composition sterilized under the same conditions as above, and the microorganism was aerobically cultured for 3 days under shaking at 30 °C, 0.25 vvm, and 70 rpm.
  • the microorganism was removed from 300 l of the culture solution through ultrafiltration membrane Microza® produced by Asahi Kasei Co., Ltd.), and the microorganism-free culture was concentrated to 6.3 l of liquid through a hollow fiber membrane (cut off molecular weight > 6000), and the resulting concentrate was divided into 900-ml aliquots and stored -20 °C.
  • 900 ml of the concentrate was used for purification of cycloisomaltooligosaccharide synthase.
  • the concentrate was thawed and then dialyzed overnight at 4 °C against 10 mM phosphate buffer, pH 7.0, containing 1 mM EDTA.
  • the dialyzate was centrifuged for removal of insolubles, and the supernatant was applied to DEAE-Sepharose CL6B column pre-equilibrated with the same buffer. After washed with the same buffer, the absorbed protein was eluted with 0-0.8 M linear gradient of NaCl.
  • the fractions with the enzyme activity, 320 ml, were combined, and ammonium sulfate was added at a final concentration of 1 M.
  • the fractions with the enzyme activity 88 ml, were collected and concentrated into about 1 ml liquid which in turn was diluted with 20 ml of 10 mM phosphate buffer, pH 7.0, containing 1 mM EDTA. This solution was applied to TSK gel DEAE 5PW column pre-equilibrated with 10 mM phosphate buffer, pH 7.0, containing 1 mM EDTA, then washed with the same buffer, and washed once with 0.15 M NaCl. The absorbed protein was eluted with 0.15-0.4 M linear gradient of NaCl.
  • the fractions with the enzyme activity 12 ml, were collected and concentrated to 0.9 ml liquid by ultrafiltration.
  • 0.3 ml aliquot of the resultant enzyme concentrate was applied to TSK gel G3000SW column pre-equilibrated with 100 mM phosphate buffer, pH 7.0, containing 10 mM EDTA and 200 mM NaCl, and was then eluted with the same buffer.
  • the remaining enzyme concentrate 0.6 ml, was divided into 0.3-ml aliquots, and each aliquot was applied to the column in the same manner as described above.
  • the fractions with the enzyme activity were collected and analyzed by SDS-PAGE. The result showed one single band, indicating that the sample is free from any impurities.
  • the purified cycloisomaltooligosaccharide synthase 12 mg, was obtained in the above procedure.
  • the physicochemical properties of the present purified enzyme were as set forth above. It was confirmed that dextran incubated with the present purified enzyme forms cycloisomaltooligosaccharides.
  • 300 l liquid culture of Bacillus sp. T-3040 was passed through a Microza® membrane for removal of the microorganism, and the microorganism-free culture was concentrated to 6.3 l liquid through a hollow fiber membrane (cut off molecular weight > 6000), and was then divided into aliquots each 900 ml for storage at -20 °C.
  • a part of the culture concentrate was mixed with 10 l aqueous solution of 100 g dextran (produced by Meito Sangyo Co. Ltd.) in 10 mM phosphate buffer, pH 6.5, and the mixture was incubated for 48 hours at 40 °C. The solution was boiled for termination of the reaction, with activated charcoal added for adsorption of unreacted dextran.
  • the weights of the resulting cycloisomaltoligosaccharides are 2.1 g cycloisomaltoheptaose, 4.6 g cycloisomaltooctaose, and 1.0 g cycloisomaltononaose.
  • the yield of the cycloisomaltooligosaccharides in total was 7.7 %.
  • HPLC analysis indicated that the purity of each oligosaccharide was 98 % or more.
  • FIG. 4 shows profiles in HPLC of the cycloisomaltooligosaccharides.
  • HPLC analysis was conducted on TSK gel Amide (a partition-adsorption chromatography column produced by Tohso Co., Ltd.).
  • A is the cycloisomaltoheptaose fraction
  • B the cycloisomaltooctaose fraction
  • C the cycloisomaltononaose fraction
  • D the standard.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Materials Engineering (AREA)
  • Polymers & Plastics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)

Claims (10)

  1. Cycloisomaltooligosaccharid, gewählt aus der Cycloisomaltoheptaose mit einer aus 7 Glukoseresten in α-1,6-Bindung aufgebauten cyclischen Struktur, Cyclolsomaltooctaose mit einer aus 8 Glukoseresten in α-1,6-Bindung aufgebauten cyclischen Struktur und Cycloisomaltononaose mit einer aus 9 Glukoseresten in α-1,6-Bindung aufgebauten cyclischen Struktur umfassenden Gruppe.
  2. Cycloisomaltooligosaccharid nach Anspruch 1, wiedergegeben durch Formel I:
    Figure imgb0007
     worin n für eine ganze Zahl von 7-9 steht.
  3. Cycloisomaltooligosaccharid nach Anspruch 2, wobei n für 7 steht.
  4. Cycloisomaltooligosaccharid nach Anspruch 2, wobei n für 8 steht.
  5. Cycloisomaltooligosaccharid nach Anspruch 2, wobei n für 9 steht.
  6. Cyclolsomaltooligosaccharid-Synthase mit den physikochemischen Eigenschaften:
    ① Wirkung auf ein Polymer aus Glukosen, die durch α-1,6-Bindungen verknüpft sind, wie Dextran, zur Bildung eines Cycloisomaltooligosaccharids durch intramolekulare Transglykosylierungsreaktion:
    ② Substratspezifität der Wirkung auf Dextran mit einer α-1,6-Bindung als eine Hauptkette, jedoch nicht auf Amylopektin und Pullulan mit einer nur teilweisen α-1,6-Bindung von Glukosen in der Struktur: und
    ③ optimaler pH in der Umgebung von pH 5,5 und Stabilitäts-pH im Bereich von pH 4,5-8,5.
  7. Verfahren zur Herstellung von Cycloisomaltooligosaccharid-Synthase, umfassend das Kultivieren in einem dextranhaltigen Medium eines zur Gattung Bacillus gehörigen Mikroorganismus, der in der Lage ist, das Enzym gemäß Anspruch 6 zu produzieren, und dann Gewinnen des Enzyms aus der Kultur.
  8. Verfahren zur Herstellung der Cycloisomaltooligosaccharide nach Anspruch 1, umfassend das Wirkenlassen des Enzyms nach Anspruch 6 auf Dextran, um Cycloisomaltooligosaccharide zu bilden.
  9. Verfahren zur Herstellung der Cycloisomaltooligosaccharide nach Anspruch 1, umfassend das Wirkenlassen einer mikroorganismusfreien Mischung, die durch Kultivieren in einem dextranhaltigen Medium eines zur Gattung Bacillus gehörigen Mikroorganismus, der in der Lage ist, das Enzym nach Anspruch 6 zu produzieren, erhalten worden ist, auf Dextran, um die Cycloisomaltooligosaccharide zu bilden.
  10. Verfahren zur Herstellung von Cycloisomaltooligosacchariden, umfassend das Kultivieren in einem α-1,6-glukanhaltigen Medium eines zur Gattung Bacillus gehörigen Mikroorganismus, der in der Lage ist, das Cycloisomaltooligosaccharid gemäß Anspruch 1 zu produzieren, und dann Gewinnen des Cycloisomaltooligosaccharids aus der Kultur.
EP93310579A 1992-12-28 1993-12-24 Cycloisomaltooligosaccharide; Enzym und Verfahren zu ihrer Herstellung, und Verfahren zur Herstellung von dem Enzym Expired - Lifetime EP0608636B1 (de)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
JP34944492A JP3075873B2 (ja) 1992-12-28 1992-12-28 新規な環状イソマルトオリゴ糖及びその製造法
JP349444/92 1992-12-28
JP05153456A JP3117328B2 (ja) 1993-06-24 1993-06-24 新規な環状イソマルトオリゴ糖合成酵素、その製造法及び環状イソマルトオリゴ糖の製造法
JP153456/93 1993-06-24

Publications (2)

Publication Number Publication Date
EP0608636A1 EP0608636A1 (de) 1994-08-03
EP0608636B1 true EP0608636B1 (de) 1997-03-05

Family

ID=26482076

Family Applications (1)

Application Number Title Priority Date Filing Date
EP93310579A Expired - Lifetime EP0608636B1 (de) 1992-12-28 1993-12-24 Cycloisomaltooligosaccharide; Enzym und Verfahren zu ihrer Herstellung, und Verfahren zur Herstellung von dem Enzym

Country Status (3)

Country Link
US (2) US5364936A (de)
EP (1) EP0608636B1 (de)
DE (1) DE69308531T2 (de)

Families Citing this family (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5686132A (en) * 1994-04-01 1997-11-11 Ezaki Glico Co., Ltd. Glucans having a cycle structure, and processes for preparing the same
JP3400868B2 (ja) * 1994-08-18 2003-04-28 独立行政法人食品総合研究所 抗う蝕剤
JP3429569B2 (ja) * 1994-08-30 2003-07-22 財団法人野田産業科学研究所 環状イソマルトオリゴ糖合成酵素遺伝子を含有するｄｎａ、組み換え体ｄｎａ、及び環状イソマルトオリゴ糖合成酵素の製造法
US6248566B1 (en) 1994-09-13 2001-06-19 Ezaki Glico Co., Ltd. Glucan having cyclic structure and method for producing the same
US5827697A (en) * 1995-03-31 1998-10-27 Ezaki Glico Co., Ltd. Process for preparing glucans having a cyclic structure
JPH11100401A (ja) * 1997-07-30 1999-04-13 Kikkoman Corp 環状オリゴ糖及びそれを含むレトロウイルス性疾患の予防または治療剤
DE19827978C2 (de) * 1998-06-24 2000-11-30 Aventis Res & Tech Gmbh & Co Verfahren zur Herstellung wasserunlöslicher alpha-1,4 Glucane
KR100875576B1 (ko) * 2001-04-27 2008-12-23 가부시끼가이샤 하야시바라 세이부쓰 가가꾸 겐꾸조 이소말토오스의 제조방법 및 용도
TWI324635B (en) 2001-10-18 2010-05-11 Hayashibara Biochem Lab Process for producing isomaltitol and uses thereof
US20040143294A1 (en) * 2003-01-22 2004-07-22 Cardia, Inc. Septal stabilization device
CN1273610C (zh) * 2004-01-15 2006-09-06 华南理工大学 一种祛除低聚糖中单糖成分的方法
KR101157230B1 (ko) * 2004-08-17 2012-06-15 삼성코닝정밀소재 주식회사 나노 기공을 포함하는 물질을 형성하기 위한 조성물
KR102026752B1 (ko) * 2014-09-22 2019-09-30 니혼 쇼꾸힌 카코 가부시키가이샤 지소화성 지속형 에너지 보급제

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
LU29750A1 (de) *
US4135977A (en) * 1974-06-20 1979-01-23 Rikagaku Kenkyusho Process for production of cyclodextrin

Also Published As

Publication number Publication date
DE69308531T2 (de) 1997-06-26
DE69308531D1 (de) 1997-04-10
EP0608636A1 (de) 1994-08-03
US5453369A (en) 1995-09-26
US5364936A (en) 1994-11-15

Similar Documents

Publication Publication Date Title
EP0327099B1 (de) Cyclomaltodextrin-Glucanotransferase, Verfahren zu ihrer Herstellung und für das Verfahren nützlicher Mikroorganismus
EP0608636A1 (de) Cycloisomaltooligosaccharide; Enzym und Verfahren zu ihrer Herstellung, und Verfahren zur Herstellung von dem Enzym
CA2107124A1 (en) Anticoagulants and processes for preparing such
WO2001002597A1 (en) Process for the preparation of the polysaccharides k4 and k5 from escherichia coli
US4338433A (en) Moranoline derivatives and process for preparation thereof
JP3490481B2 (ja) オリゴ糖の製造方法
US5861289A (en) Heat-resistant β-galactosyltransferase, its production process and its use
US5153128A (en) Heat-resistant β-galactosyltransferase, its production process and its use
JP3075873B2 (ja) 新規な環状イソマルトオリゴ糖及びその製造法
FI110518B (fi) Levaanisakkaraasientsyymi, menetelmä sen tuottamiseksi, sitä tuottavia mikro-organismeja ja sitä sisältävä koostumus
JP3117328B2 (ja) 新規な環状イソマルトオリゴ糖合成酵素、その製造法及び環状イソマルトオリゴ糖の製造法
JP2000125857A (ja) α−Ｌ−フコシダーゼ、その製造方法、菌株および用途
JP3673302B2 (ja) 分岐サイクロイソマルトヘプタオース及びその製造法
JPH04200386A (ja) β―フラクトフラノシダーゼ及びその製造方法
JPH06133791A (ja) 低分子デキストラン
JP3556704B2 (ja) β−ガラクトシダーゼ
EP0193608A1 (de) Gif-2 und dessen herstellung
JP3682931B2 (ja) ガラクトシル−マルトオリゴ糖誘導体の製造方法
JPH0437719B2 (de)
KR830001418B1 (ko) 모라노린 유도체의 제조방법
KR840000893B1 (ko) 모라노린 유도체의 제조방법
JP2819310B2 (ja) 新規エキソ―β―Ｄ―グルコサミニダーゼ及びその製造法
JPH06225755A (ja) デキストランスクラーゼ生産性新規微生物及びこの新規微生物を用いるデキストランスクラーゼの生産方法
EP0577665A1 (de) Antikoagulantien und verfahren zu deren herstellung
KR20050027440A (ko) 베타-프럭토실-엘-아스코르브산 및 그의 제조방법

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): DE FR GB NL SE

17P Request for examination filed

Effective date: 19941017

GRAG Despatch of communication of intention to grant

Free format text: ORIGINAL CODE: EPIDOS AGRA

17Q First examination report despatched

Effective date: 19960410

GRAH Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOS IGRA

GRAH Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOS IGRA

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): DE FR GB NL SE

ET Fr: translation filed
REF Corresponds to:

Ref document number: 69308531

Country of ref document: DE

Date of ref document: 19970410

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed
PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20011227

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 20011228

Year of fee payment: 9

REG Reference to a national code

Ref country code: GB

Ref legal event code: IF02

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GB

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20021224

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NL

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20030701

GBPC Gb: european patent ceased through non-payment of renewal fee

Effective date: 20021224

NLV4 Nl: lapsed or anulled due to non-payment of the annual fee

Effective date: 20030701

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20101224

Year of fee payment: 18

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: SE

Payment date: 20101213

Year of fee payment: 18

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20101222

Year of fee payment: 18

REG Reference to a national code

Ref country code: SE

Ref legal event code: EUG

REG Reference to a national code

Ref country code: FR

Ref legal event code: ST

Effective date: 20120831

REG Reference to a national code

Ref country code: DE

Ref legal event code: R119

Ref document number: 69308531

Country of ref document: DE

Effective date: 20120703

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20120703

Ref country code: SE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20111225

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: FR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20120102